Primitive hematopoietic progenitors within mobilized blood are spared by uncontrolled rate freezing

D. Cilloni, D. Garau, E. Regazzi, G. Sammarelli, B. Savoldo, C. Caramatti, L. Mangoni, V. Rizzoli, C. Carlo-Stella

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Uncontrolled-rate freezing techniques represent an attractive alternative to controlled-rate cryopreservation procedures which are time-consuming and require high-level technical expertise. In this study, we report our experience using uncontrolled-rate cryopreservation and mechanical freezer storage at -140°C. Twenty-eight PBPC samples (10 cryovials, 18 freezing bags) from 23 patients were cryopreserved in a cryoprotectant solution composed of phosphate-buffered saline (80%, v/v) supplemented with human serum albumin (10%, v/v) and dimethylsulfoxide (10%, v/v). The cryopreservation procedure required on average 1.5 h. The mean (± s.e.m.) storage time of cryovials and bags was 344 ± 40 and 299 ± 57 days, respectively. Although cell thawing was associated with a statistically significant reduction of the absolute number of nucleated cells (vials: 0.3 x 109 vs 0.2 x 109, P ≤ 0.02; bags: 14 x 109 vs 11 x 109, P ≤ 0.0003), the growth of committed progenitors was substantially unaffected by the freezing-thawing procedure, with mean recoveries of CFU-Mix, BFU-E, and CFU-GM ranging from 60 ± 29% to 134 ± 15%. Mean recoveries of LTC-IC from cryovials and bags were 262 ± 101% and 155 ± 27% (P ≤ 0.2), respectively. In 14 out of 23 patients who underwent high-dose chemotherapy and PBPC reinfusion, the pre- and post-freezing absolute numbers of hematopoietic progenitors cryopreserved in bags were compared. A significant reduction was detected for CFU-Mix (11 vs 7.4 x 105), but no significant loss of BFU-E (180 vs 150 x 105), CFU-GM (400 vs 290 x 105) and LTC-IC (15 vs 16 x 105) could be demonstrated. When these patients were reinfused with uncontrolled-rate cryopreserved PBPC, the mean number of days to reach 1 x 109/l white blood cells and 50 x 109/l platelets were 9 and 13, respectively. In conclusion, the procedure described here is characterized by short execution time, allows a substantial recovery of primitive and committed progenitors and is associated with prompt hematopoietic recovery following myeloablative therapy even after long-term storage.

Original languageEnglish
Pages (from-to)497-503
Number of pages7
JournalBone Marrow Transplantation
Volume23
Issue number5
Publication statusPublished - 1999

Fingerprint

Freezing
Cryopreservation
Granulocyte-Macrophage Progenitor Cells
Erythroid Precursor Cells
Professional Competence
Dimethyl Sulfoxide
Serum Albumin
Leukocytes
Blood Platelets
Cell Count
Phosphates
Drug Therapy
Growth
Therapeutics

Keywords

  • Blood stem cell transplantation
  • Colony-forming cells
  • Cryopreservation
  • LTC-IC
  • PBPC

ASJC Scopus subject areas

  • Hematology
  • Transplantation

Cite this

Cilloni, D., Garau, D., Regazzi, E., Sammarelli, G., Savoldo, B., Caramatti, C., ... Carlo-Stella, C. (1999). Primitive hematopoietic progenitors within mobilized blood are spared by uncontrolled rate freezing. Bone Marrow Transplantation, 23(5), 497-503.

Primitive hematopoietic progenitors within mobilized blood are spared by uncontrolled rate freezing. / Cilloni, D.; Garau, D.; Regazzi, E.; Sammarelli, G.; Savoldo, B.; Caramatti, C.; Mangoni, L.; Rizzoli, V.; Carlo-Stella, C.

In: Bone Marrow Transplantation, Vol. 23, No. 5, 1999, p. 497-503.

Research output: Contribution to journalArticle

Cilloni, D, Garau, D, Regazzi, E, Sammarelli, G, Savoldo, B, Caramatti, C, Mangoni, L, Rizzoli, V & Carlo-Stella, C 1999, 'Primitive hematopoietic progenitors within mobilized blood are spared by uncontrolled rate freezing', Bone Marrow Transplantation, vol. 23, no. 5, pp. 497-503.
Cilloni D, Garau D, Regazzi E, Sammarelli G, Savoldo B, Caramatti C et al. Primitive hematopoietic progenitors within mobilized blood are spared by uncontrolled rate freezing. Bone Marrow Transplantation. 1999;23(5):497-503.
Cilloni, D. ; Garau, D. ; Regazzi, E. ; Sammarelli, G. ; Savoldo, B. ; Caramatti, C. ; Mangoni, L. ; Rizzoli, V. ; Carlo-Stella, C. / Primitive hematopoietic progenitors within mobilized blood are spared by uncontrolled rate freezing. In: Bone Marrow Transplantation. 1999 ; Vol. 23, No. 5. pp. 497-503.
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abstract = "Uncontrolled-rate freezing techniques represent an attractive alternative to controlled-rate cryopreservation procedures which are time-consuming and require high-level technical expertise. In this study, we report our experience using uncontrolled-rate cryopreservation and mechanical freezer storage at -140°C. Twenty-eight PBPC samples (10 cryovials, 18 freezing bags) from 23 patients were cryopreserved in a cryoprotectant solution composed of phosphate-buffered saline (80{\%}, v/v) supplemented with human serum albumin (10{\%}, v/v) and dimethylsulfoxide (10{\%}, v/v). The cryopreservation procedure required on average 1.5 h. The mean (± s.e.m.) storage time of cryovials and bags was 344 ± 40 and 299 ± 57 days, respectively. Although cell thawing was associated with a statistically significant reduction of the absolute number of nucleated cells (vials: 0.3 x 109 vs 0.2 x 109, P ≤ 0.02; bags: 14 x 109 vs 11 x 109, P ≤ 0.0003), the growth of committed progenitors was substantially unaffected by the freezing-thawing procedure, with mean recoveries of CFU-Mix, BFU-E, and CFU-GM ranging from 60 ± 29{\%} to 134 ± 15{\%}. Mean recoveries of LTC-IC from cryovials and bags were 262 ± 101{\%} and 155 ± 27{\%} (P ≤ 0.2), respectively. In 14 out of 23 patients who underwent high-dose chemotherapy and PBPC reinfusion, the pre- and post-freezing absolute numbers of hematopoietic progenitors cryopreserved in bags were compared. A significant reduction was detected for CFU-Mix (11 vs 7.4 x 105), but no significant loss of BFU-E (180 vs 150 x 105), CFU-GM (400 vs 290 x 105) and LTC-IC (15 vs 16 x 105) could be demonstrated. When these patients were reinfused with uncontrolled-rate cryopreserved PBPC, the mean number of days to reach 1 x 109/l white blood cells and 50 x 109/l platelets were 9 and 13, respectively. In conclusion, the procedure described here is characterized by short execution time, allows a substantial recovery of primitive and committed progenitors and is associated with prompt hematopoietic recovery following myeloablative therapy even after long-term storage.",
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AU - Carlo-Stella, C.

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